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Nan Fang Yi Ke Da Xue Xue Bao ; 36(11): 1482-1488, 2016 Nov 20.
Artigo em Chinês | MEDLINE | ID: mdl-27881337

RESUMO

OBJECTIVE: To explore the inhibitory effect of migration-inducing gene 7 (Mig-7) gene silencing induced by retroviral-mediated small hairpin RNA (shRNA) on vasculogenic mimicry (VM), invasion and metastasis of human hepatocellular carcinoma (HCC) cells in vitro. METHODS: Two target sequences (Mig-7 shRNA-1 and Mig-7 shRNA-2) and one negative control sequence (Mig-7 shRNA-N) were synthesized. The recombinant retroviral vectors carrying Mig-7 shRNA were constructed, and HCC cell line MHCC-97H were transfected with Mig-7 shRNA-1, Mig-7 shRNA-2, Mig-7 shRNA-N, or the empty vector, or treated with 125 µg/mL recombinant human endostatin (ES). Mig-7 expression in the treated cells was detected using semi-quantitative PCR and Western blotting. The inhibitory effect of Mig-7 silencing on VM formation was investigated in a 3-dimensional cell culture system; the changes in cell adhesion, invasion and migration were assessed with intercellular adhesion assay, Transwell invasion assay and Transwell migration assay, respectively. RESULTS: The expression of Mig-7 at both mRNA and protein levels decreased significantly, VM formation, invasion and metastasis were suppressed, while intercellular adhesion increased significantly in MHCC-97H cells in Mig-7 shRNA-1 and Mig-7 shRNA-2 groups (P<0.05); such changes were not observed in cells transfected with Mig-7 shRNA-N or the empty vector, nor in cells treated with ES. CONCLUSIONS: Mig-7 silencing by retroviral-mediated shRNA significantly inhibits VM formation, invasion and metastasis and increases the intercellular adhesion of the HCC cells, while ES does not have such inhibitory effects.


Assuntos
Carcinoma Hepatocelular/patologia , Inativação Gênica , Neoplasias Hepáticas/patologia , Proteínas de Neoplasias/genética , RNA Interferente Pequeno , Carcinoma Hepatocelular/genética , Linhagem Celular Tumoral , Endostatinas , Vetores Genéticos , Humanos , Neoplasias Hepáticas/genética , RNA Mensageiro , Retroviridae , Transfecção
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